Publication

Combining 2-DE immunoblots and mass spectrometry to identify putative soybean (Glycine max) allergens

Journal Paper/Review - Apr 16, 2018

Units
PubMed
Doi

Citation
Lu M, Jin Y, Cerny R, Ballmer-Weber B, Goodman R. Combining 2-DE immunoblots and mass spectrometry to identify putative soybean (Glycine max) allergens. Food Chem Toxicol 2018; 116:207-215.
Type
Journal Paper/Review (English)
Journal
Food Chem Toxicol 2018; 116
Publication Date
Apr 16, 2018
Issn Electronic
1873-6351
Pages
207-215
Brief description/objective

Soybean is recognized as a commonly allergenic food, but the identity of important allergens is not well studied. Recently, some global regulatory agencies started requiring quantitative analysis of individual allergens, including unproven allergens, as part of the risk assessment for genetically engineered (GE) soybeans. We sought to identify soybean proteins that bind IgE from any of 10 individual soybean-sensitized subjects. Soybean IgE binding proteins were identified by 2-DE immunoblots using sera from four soy-allergic and plasma from six soy-sensitized human subjects. Corresponding spots were excised from stained gels, digested, and analyzed using a quadrupole TOF Synapt G2-S tandem mass spectrometer. Results showed the major IgE binding proteins were subunits of either β-conglycinin (Gly m 5) or glycinin (Gly m 6). Soybean Kunitz trypsin inhibitor (SKTI) was a significant IgE binding protein for four subjects. Soybean agglutinin, seed biotinylated protein (SBP) of 65 kDa, late embryogenesis protein (LEP), and sucrose-binding protein were identified as IgE binding only for soy-sensitized subjects. We conclude that the major soybean allergens are isoforms of Gly m 5, Gly m 6, and possibly SKTI and that requirements for quantitative measurement of proteins that are not clear allergens is not relevant to safety.